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A novel phosphate-regulated expression vector in Escherichia coli

dc.contributor.authorSu, Ti-Zhien_US
dc.contributor.authorSchweizer, Herbert P.en_US
dc.contributor.authorOxender, Dale L.en_US
dc.date.accessioned2006-04-10T13:43:35Z
dc.date.available2006-04-10T13:43:35Z
dc.date.issued1990-05-31en_US
dc.identifier.citationSu, Ti-Zhi, Schweizer, Herbert, Oxender, Dale L. (1990/05/31)."A novel phosphate-regulated expression vector in Escherichia coli." Gene 90(1): 129-133. <http://hdl.handle.net/2027.42/28559>en_US
dc.identifier.urihttp://www.sciencedirect.com/science/article/B6T39-47GH971-M/2/5764f4602ab23d6426fbf5c41dd54305en_US
dc.identifier.urihttps://hdl.handle.net/2027.42/28559
dc.identifier.urihttp://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=2379833&dopt=citationen_US
dc.description.abstractThe ugp promoter (pugp) responsible for expression of the binding-protein-dependent sn-glycerol-3-phosphate transport system in Escherichia coli was cloned into a small multicopy plasmid pTER5, a derivative of pBR322, between the transcription terminators rpoCt and tL1. The resulting expression vector, pPH3, permits convenient insertion of structural genes containing their own translational-initiation regions, into the multiple-cloning site derived from the pUC19 plasmid. The efficiency and regulatory properties of pugp were measured using xylE and lacZ as reporter genes, which code for the corresponding enzymes catechol-2,3-dioxygenase (C23O) and [beta]-galactosidase ([beta]Gal), respectively. Enzyme activities were virtually completely repressed in the presencee of excess inorganic phosphates (Pi) and high concentrations of glucose. Maximal induction was observed at limiting Pi (pugp -directed [beta]Gal synthesis was approx. 80% of that directed by directed by strong ptac. When the xylE gene was maximally expressed, the induced enzyme constituted approx. 50% of total cellular protein as judged by laser densitometry following sodium dodecyl sulfate-polyacrylamide-gel electrophoresis. These results suggest the usefulness of the pugp in expression vectors for strong, but controlled, expression of cloned genes in E. coli. This Pi controlled vector can be adapted to large-scale fermentation by using Pi-limiting growth conditions.en_US
dc.format.extent367422 bytes
dc.format.extent3118 bytes
dc.format.mimetypeapplication/pdf
dc.format.mimetypetext/plain
dc.language.isoen_US
dc.publisherElsevieren_US
dc.titleA novel phosphate-regulated expression vector in Escherichia colien_US
dc.typeArticleen_US
dc.rights.robotsIndexNoFollowen_US
dc.subject.hlbsecondlevelMolecular, Cellular and Developmental Biologyen_US
dc.subject.hlbsecondlevelGeneticsen_US
dc.subject.hlbsecondlevelBiological Chemistryen_US
dc.subject.hlbtoplevelScienceen_US
dc.subject.hlbtoplevelHealth Sciencesen_US
dc.description.peerreviewedPeer Revieweden_US
dc.contributor.affiliationumDepartment of Biological Chemistry, University of Michigan, Ann Arbor, MI 48109-0606, U.S.A.en_US
dc.contributor.affiliationumDepartment of Biological Chemistry, University of Michigan, Ann Arbor, MI 48109-0606, U.S.A.en_US
dc.contributor.affiliationotherDepartment of Microbiology, University of Calgary, Calgary, Alberta, Canada T2N 4N1en_US
dc.identifier.pmid2379833en_US
dc.description.bitstreamurlhttp://deepblue.lib.umich.edu/bitstream/2027.42/28559/1/0000361.pdfen_US
dc.identifier.doihttp://dx.doi.org/10.1016/0378-1119(90)90448-Zen_US
dc.identifier.sourceGeneen_US
dc.owningcollnameInterdisciplinary and Peer-Reviewed


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